王婷,李煜江,叶雅雯,等.运动对2型糖尿病小鼠心肌中促纤维化基因的影响[J].中华物理医学与康复杂志,2025,47(6):481-486
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| 运动对2型糖尿病小鼠心肌中促纤维化基因的影响 |
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| DOI：10.3760/cma.j.cn421666-20240106-00014 |
| 中文关键词: 运动 糖尿病 心肌病 miR-344g-5p 纤维化 |
| 英文关键词: Exercise Diabetes Cardiomyopathy miR-344g-5p genes Fibrosis |
| 基金项目:浙江省自然科学基金（LY22H020002）；2022年浙江省卫生健康科技计划(2022KY900)；国家自然科学基金（82272603） |
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| 中文摘要: |
| 目的 观察运动调节miR-344g-5p对糖尿病小鼠心肌中促纤维化基因母体信号蛋白同源物（SMAD）和转化生长因子-β（TGF-β）的影响。 方法 选取雄性C57BL/6小鼠(6～8周龄)24只，采用随机数字表法分为对照组12只和2型糖尿病组12只，按运动情况再将2组小鼠各分为静养和运动2个亚组（即静养对照组、运动对照组、静养2型糖尿病组和运动2型糖尿病组），每个亚组6只小鼠。入组后，对照组的饲料为普通饲料，2型糖尿病组的饲料为高脂饲料，入组12周后，2型糖尿病组采用腹腔注射链脲佐菌素诱导2型糖尿病模型。入组14周后，运动对照组和运动2型糖尿病组开始游泳训练，每日1次，每次40 min，每周训练5 d，连续训练8周。入组22周后，应用小动物超声仪检测4个亚组小鼠的心脏功能，检测结束后处死，取心肌组织包埋切片用天狼猩红固绿染色，应用qRT-PCR检测心肌中miR-344g-5p表达；采用蛋白免疫印迹检测磷酸化SMAD3(p-SMAD3)和TGF-β的蛋白表达；检索Target Scan数据库，进一步分析miR-344g-5p与促纤维化基因SMAD3、TGFBR2、COL1A2和COL12A1等是否存在预测靶点，并确定基因调控的相关性。 结果 入组22周后，静养2型糖尿病组的EF和E/A比值分别为（57.5±4.1）%和（1.4±0.3），均显著低于静养对照组、运动对照组和运动2型糖尿病，差异均有统计学意义（P＜0.05）。入组22周后，静养2型糖尿病组的心肌胶原纤维较静养对照组和运动2型糖尿病组均显著增加，差异均有统计学意义(P＜0.05)。入组22周后，运动2型糖尿病组心肌中miR-344g-5p的表达较静养2型糖尿病组显著增加（P＜0.05）。入组22周后，静养2型糖尿病组心肌中p-SMAD3和TGF-β的表达均显著高于静养对照组和运动2型糖尿病组（P＜0.05）。在Target Scan数据库中分析miR-344g-5p的预测靶作用基因，结果发现，miR-344g-5p与SMAD3、TGFBR2、COL1A2和COL12A1等多个促纤维化基因存在潜在结合位点，结合TGF-β和p-SMAD蛋白表达在运动2型糖尿病组降低的结果推测，miR-344g-5p可能抑制这些基因转录后的翻译过程。 结论 运动可通过增加2型糖尿病小鼠心肌中miR-344g-5p的表达，靶向抑制p-SMAD3和TGF-β蛋白的表达，减少糖尿病心肌纤维化，从而促进糖尿病心肌病的康复。 |
| 英文摘要: |
| Objective To observe the effect of exercise-regulated miR-344g-5p on the fibrosis-related SMAD genes and transforming growth factor beta (TGF-β) in the myocardia of mice modeling diabetes. Methods Twenty-four male C57BL/6 mice (6-8 weeks old) were randomly divided into a control group (n=12) and a type 2 diabetes group (n=12). Each group was further divided into two subgroups based on exercise status to form a sedentary control group, an exercise control, a sedentary type 2 diabetes group and an exercise type 2 diabetes group with six mice in each subgroup. The control groups were fed a normal diet, while the type 2 diabetes groups were on a high-fat diet for 12 weeks. Type 2 diabetes was then induced by intraperitoneal injection of streptozotocin. Two weeks later, the exercise groups began 40 minutes of daily swimming training, five days a week for eight consecutive weeks. Right after that, their cardiac function was measured using a small animal ultrasound system and the derived ejection fraction (EF) and the maximal early (E) and late (A) transmitral velocities ratio (E/A ratio) in diastole. They were then sacrificed and myocardial tissue was resected and stained with Sirius red. The expression of miR-344g-5p in the myocardium was detected using quantitative polymerase chain reactions. The expression of phosphorylated SMAD3 (p-SMAD3) and TGF-β were assessed using western blotting. The Target Scan database was exploited to analyze whether there were predicted targets of miR-344g-5p and pro-fibrotic genes such as SMAD3, TGFBR2, COL1A2 and COL12A1, and to determine any correlations in the gene regulation. Results After 22 weeks, the EF and E/A ratio in the sedentary type 2 diabetes group were (57.5±4.1)% and (1.4±0.3), respectively, both significantly lower than in the other groups. Myocardial collagen fibers in the sedentary type 2 diabetes group were significantly more abundant than in the sedentary control and exercise type 2 diabetes groups. And miR-344g-5p expression in the myocardia of the exercise type 2 diabetes group was significantly greater than that in the sedentary type 2 diabetes group. The expression of p-SMAD3 and TGF-β in the myocardia of the sedentary type 2 diabetes group was significantly higher than in the sedentary control and exercise type 2 diabetes groups. Target Scan analysis revealed that miR-344g-5p had potential binding sites with several fibrosis-related genes such as SMAD3, TGFBR2, COL1A2, and COL12A1. Based on the reduction in TGF-β and p-SMAD protein expression in the exercise type 2 diabetes group, it was hypothesized that miR-344g-5p may inhibit the post-transcriptional processes of those genes. Conclusions Exercise promotes the recovery of diabetic cardiomyopathy by upregulating myocardial miR-344g-5p expression, which subsequently targets and suppresses p-SMAD3 and TGF-β protein expression, thereby reducing diabetic myocardial fibrosis. |
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